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Nature is considered a prolific source of diverse biologically active chemotypes. Medicinal plants have historically attracted the most attention and have led to the discovery of various drugs ranging from anticancer to anti-infectious agents. However, a vast array of bioactive metabolites have also been discovered from marine organisms, fungi and bacteria. While most reviews have focused on the characteristics of the chemical backbones of natural products (NPs), few have tried to provide an overview of their origins (the living organisms in which they are produced), chemical classes, and biological activities. This review discusses the current knowledge on NP diversity by focusing on the Dictionary of Natural Products® (DNP). We datamined the 300,000 NPs covered by the DNP to reveal relevant, albeit dormant, knowledge about NP diversity. This holistic picture of NPs allows us to discuss the most abundant biological sources of NPs investigated in relation to their chemical features and biological activities. In a nutshell, a large part of NPs originated from plants (67%), especially from the Compositae and Leguminosae families. Among all kingdoms, NPs isolated from Streptomyces spp. were largely represented, while terpenoids and alkaloids were the two most represented chemical classes. Out of all NPs documented, only 3,882 were reported to be bioactive (1,163 from plants and 1,006 from bacteria), with antibacterial, antibiotics, and antineoplastic agents being the most frequent therapeutic classes. In this paper, we also address the advantages and limitations of NP research from a pharmaceutical industry perspective. This work will provide useful insights and guidance to researchers involved in drug discovery from NPs.

International audience; Container-content interactions are common in the food and pharmaceutical industries. However, these studies are more complicated in the cosmetic industry, and it is necessary to ensure consumer safety. The objective of this work was to develop a strategy for the toxicological evaluation of leachables for cosmetic packagings. Eleven common plastic packagings were selected to evaluate interactions with 5 simulants (acidic, alkaline and neutral water, 30% and 96% ethanol) chosen to mimic cosmetics behavior. A GC-MS method was developed to screen for 12 non-intentionally added substances of particular concern: 10 phthalates, bisphenol A and distearyl thiodipropionate (European Pharmacopoeia plastic additive 17). Results were analyzed using a toxicological procedure established for this study. Some phthalates and bisphenol A were detected in several samples, but only one contaminant, diisobutyl phthalate (DiBP), was found to be above the set concentration threshold. Using toxicological data, this concentration was found to be safe for users. 96% ethanol appeared to be the strongest simulant in term of extraction, with a maximum concentration of 491 μg/L for DiBP in a 100% styrene-acrylonitrile copolymer packaging. In water simulants, less contaminants were extracted, with concentrations under 20 μg/L.

International audience; Skin aging is a complex biological process mixing intrinsic and extrinsic factors, such as sun exposure. At the molecular level, skin aging affects in particular the extracellular matrix proteins. Materials and Methods: Using Raman imaging, which is a nondestructive approach appropriate for studying biological samples, we analyzed how aging modifies the matrix proteins of the papillary and reticular dermis. Biopsies from the buttock and dorsal forearm of volunteers younger than 30 and older than 60 were analyzed in order to identify chronological and photoaging processes. Analyses were performed on skin section, and Raman spectra were acquired separately on the different dermal layers. Results: We observed differences in dermal matrix structure and hydration state with skin aging. Chronological aging alters in particular the collagen of the papillary dermis, while photoaging causes a decrease in collagen stability by altering proline and hydroxyproline residues in the reticular dermis. Moreover, chronological aging alters glycosaminoglycan content in both dermal compartments. Conclusion: Alterations of the papillary and reticular dermal matrix structures during photo-and chronological aging were clearly depicted by Raman spectroscopy.

International audience; Objective Our main objective was to compare Cutibacterium acnes (C. acnes) skin colonisation in patients with mild to moderate acne versus healthy controls and secondly, to evaluate a Myrtacine® -based cream on C. acnes total population and antibioresistant Cutibacteria in patients with acne.Methods In 60 acne patients (Global Acne Severity Scale, GEA grades 2–3), of mean age 20 [15–30] years and in 24 age-and sex-matched healthy controls, forehead strips samplings were performed for microbiological analysis of come-dones by colony forming unit (CFU) counts of global C. acnes and erythromycin (EryR) or clindamycin-resistant (ClnR) populations of Cutibacterium and determination of phylotypes by MALTI-TOF. Clinical evaluations of acne patients (GEA, lesion count, porphyrin fluorescence) were performed at baseline and after 56 days of twice-daily application of a Myrtacine® -based cream.Results We first showed (i) high and similar levels of C. acnes colonisation in superficial pilosebaceous follicles and detection of EryR and ClnR strains in both acne and control groups; (ii) different repartition of phylotypes in acne patients versus healthy control, with a predominance of phylotype IA in acne patients and a link between phylotype IA and erythromycin resistance. Besides, after treatment with the Myrtacine® -based cream in acne patients, there was no change in C. acnes total load, but a significant decrease of EryR Cutibacteria, reduced porphyrin production by C. acnes, a decrease in acne severity (GEA), associated with reduced retentional and inflammatory lesions.Conclusion Cutibacterium acnes colonisation was not significantly different in acne versus control groups. Phylotype IA was predominant in acne patient and in EryR C. acnes. A Myrtacine® -based cream significantly reduced the level of EryR Cutibacteria in vivo and improved acne lesions.

International audience; Abstract Background/purpose: The purpose of this clinical study was to identify dielectric markers to complete a previous thermal and vibrational study on the molecular and organizational changes in human dermis during intrinsic and extrinsic aging. Methods: Sun-exposed and non-exposed skin biopsies were collected from 28 women devised in two groups (20-30 and ≥60 years old). The dielectric relaxation modes associated with localized and delocalized dynamics in the fresh and dehydrated state were determined by the Thermostimulated currents technique (TSC). Results: Intrinsic and extrinsic aging induced significant evolution of some of the dielectric parameters of localized and delocalized dynamics of human skin. With photo-aging, freezable water forms a segregated phase in dermis and its dynamics is close to free water, what evidences the major role of extrinsic aging on water organization in human skin. Moreover, TSC indicators highlight the restriction of localized mobility with intrinsic aging due to glycation, and the cumulative effect of chronological aging and photo-exposition on the molecular mobility of the main structural proteins of the dermis at the mesoscopic scale. Conclusion: TSC is a well-suited technique to scan the molecular mobility of human skin. It can be uses as a relevant complement of vibrational and thermal characterization to follow human skin modifications with intrinsic and extrinsic aging.

This paper deals with the characterization and classification of reflectance confocal microscopy images of human skin. The aim is to identify and characterize the lentigo, a phenomenon that originates at the dermo-epidermic junction of the skin. High resolution confocal images are acquired at different skin depths and are analyzed for each depth. Histograms of pixel intensities associated with a given depth are determined, showing that the generalized gamma distribution (GGD) is a good statistical model for confocal images. A GGD is parameterized by translation, scale and shape parameters. These parameters are estimated using a new estimation method based on a natural gradient descent showing fast convergence properties with respect to state-of-the-art estimation methods. The resulting parameter estimates can be used to classify clinical images of healthy and lentigo patients. The obtained results show that the scale and shape parameters are good features to identify and characterize the presence of lentigo in skin tissues., Comment: 24 pages, 16 figures

International audience; Detecting skin lentigo in reflectance confocal microscopy images is an important and challenging problem. This imaging modality has not yet been widely investigated for this problem and there are a few automatic processing techniques. They are mostly based on machine learning approaches and rely on numerous classical image features that lead to high computational costs given the very large resolution of these images. This paper presents a detection method with very low computational complexity that is able to identify the skin depth at which the lentigo can be detected. The proposed method performs multiresolution decomposition of the image obtained at each skin depth. The distribution of image pixels at a given depth can be approximated accurately by a generalized Gaussian distribution whose parameters depend on the decomposition scale, resulting in a very-low-dimension parameter space. SVM classifiers are then investigated to classify the scale parameter of this distribution allowing real-time detection of lentigo. The method is applied to 45 healthy and lentigo patients from a clinical study, where sensitivity of 81.4% and specificity of 83.3% are achieved. Our results show that lentigo is identifiable at depths between 50μm and 60μm, corresponding to the average location of the the dermoepidermal junction. This result is in agreement with the clinical practices that characterize the lentigo by assessing the disorganization of the dermoepidermal junction.

International audience; Background/purpose : The purpose of this clinical study was to identify suitable biomarkers for a better understanding of the molecular and organizational changes in human dermis during intrinsic and extrinsic ageing. Methods : Sun-exposed and non-exposed skin biopsies were collected from twenty-eight women devised in two groups (20-30 and ≥60 years old). The hydric organization and thermal transitions were determined by Differential Scanning Calorimetry (DSC). Fourier Transform Infrared spectroscopy (FTIR) was used to identify the absorption bands of the dermis and to quantify the different absorbance ratio. Results : The amounts of total, freezable and unfreezable water were determined. A significant increasing amount of freezable water is evidenced in sun-exposed area skin of aged group compared with young group (P=.0126). Another significant effect of extrinsic ageing (P=.0489) is the drastic decrease of fibrillary collagen, the main protein component of dermis. The only significant effect of intrinsic ageing (P=.0184) is an increase of the heat-stable fraction of collagens in dermis. Conclusion : DSC and FTIR are well-suited techniques to characterize human skin, giving accurate results with a high reproducibility. The combination of these techniques is useful for a better understanding of human skin modifications with intrinsic and extrinsic ageing.

International audience; For a better understanding of the molecular and organizational changes in human dermis, biophysical methods were tested. The aim of this study was to find suitable and reproducible biomarkers for further clinical studies on intrinsic and extrinsic aging of dermis. Thermoporometry, hydric organization and thermal transitions of fresh and frozen skins were determined by differential scanning calorimetry (DSC). Fourier transform infrared spectroscopy (FTIR) was used to identify the absorption bands of the dermis especially in the 1800–1000 cm−1 zone and to discriminate between the different secondary structures of proteins. A widening of the pore size distribution is evidenced with freezing, but there is no significant difference between the hydric organization and the endothermic collagen denaturation of fresh and frozen skins. The global FTIR spectra and the second derivative spectra in the scanned zone are also identical in fresh and frozen dermis, validating the storage protocol. DSC and FTIR are well-suited techniques to characterize human skin, giving accurate results with high reproducibility. The acquisition of thermal and vibrational biomarkers of the skin at the mesoscale and nanoscale contributes to its better knowledge and is promising for further studies on skin aging.

International audience; The human skin microbiota is quantitatively dominated by Gram-positive bacteria, detected by both culture and metagenomics. However, metagenomics revealed a huge variety of Gram-negative taxa generally considered from environmental origin. For species affiliation of bacteria in skin microbiota, clones of 16S rRNA gene and colonies growing on diverse culture media were analyzed. Species-level identification was achieved for 81% of both clones and colonies. Fifty species distributed in 26 genera were identified by culture, mostly belonging to Actinobacteria and Firmicutes, while 45 species-level operational taxonomic units distributed in 30 genera were detected by sequencing, with a high diversity of Proteobacteria. This mixed approach allowed the detection of 100% of the genera forming the known core skin Gram-negative microbiota and 43% of the known diversity of Gram-negative genera in human skin. The orphan genera represented 50% of the current skin pan-microbiota. Improved culture conditions allowed the isolation of Roseomonas mucosa, Aurantimonas altamirensis and Agrobacterium tumefaciens strains from healthy skin. For proteobacterial species previously described in the environment, we proposed the existence of skin-specific ecotypes, which might play a role in the fine-tuning of skin homeostasis and opportunistic infections but also act as a shuttle between environmental and human microbial communities. Therefore, skin-associated proteobacteria deserve to be considered in the One-Health concept connecting human health to the health of animals and the environment.

Potentiometry and cyclic voltammetry were proposed as simple, reliable and non invasive methods for the simultaneous determination of pH and antioxidant properties of skin. Experiments were performed with microelectrodes just deposited on skin surface without any gel or water added. pH was measured by means of the zero current potential of a tungsten W/WO3 sensor. A nerstian response was recorded in pH range 4 to 6 corresponding to the normal skin pH values. The global antioxidant capacity was deduced from the anodic charge passed during the plotting of cyclic voltammograms on platinum or gold microelectrodes. Comparing the half wave or peak potentials of these curves with those recorded for experiments performed in aqueous solution, the main hydrophilic antioxidants species were detected, i.e. ascorbic acid, uric acid and glutathione. This relatively easy-to-use analytical method made it possible to follow in real time the efficiency of topic treatment as well as to study the influence of oxidative stress.

In this work, a study about solute continuous extraction from vegetable matrix is undertaken. An industrial apparatus is used and the problems are to identify information to be acquired in order to process implementation. To this end, a batch study was carried out to obtain the extraction kinetics, as well as a study of the matter behaviour under pressure and a study of raw plant transport. Finally, to couple these elements makes it possible to represent the extraction yields according to time in the apparatus. A methodology for the change from batch to continuous extractor based on laboratory tests is proposed.

BackgroundRecent works present evidence of Propionibacterium acnes growing as a biofilm in cutaneous follicles. This formation of clusters is now considered as an explanation for the in vivo resistance of P. acnes to the main antimicrobials prescribed in acne vulgaris.PurposeOur objective was to explore this hypothesis and propose a new therapeutic approach focusing on anti-biofilm activity of Myrtacine® New Generation (Mediterranean Myrtle extract–Botanical Expertise P. Fabre) alone or combined with antibiotics.Methods/ResultsUsing in vitro models able to promote the growth of adhered bacteria, the loss of sensitivity of P. acnes biofilms (48 h) towards erythromycin and clindamycin was checked considering either sensitive or resistant strains. In the same time, the activity of Myrtacine® New Generation against biofilm formation and mature biofilm (48 h) was evaluated. Using a dynamic model of biofilm formation, we noted an inhibition of biofilm formation (addition of Myrtacine® New Generation at T 0) and a significant effect on mature biofilm (48 h) for 5 min of contact. This effect was also checked using the static model of biofilm formation for Myrtacine® New Generation concentrations ranging from 0.03% to 0.0001%. A significant, dose-dependent anti-biofilm effect was observed and notable even at a concentration lower than the active concentration on planktonic cells, i.e. 0.001%. Finally, the interest of the combination of Myrtacine® New Generation with antibiotics was explored. An enhanced efficacy was noted when erythromycin (1000 mg/l) or clindamycin (500 mg/l) was added to 0.001% Myrtacine®, leading to significant differences in comparison to each compound used alone.ConclusionThe efficiency of Myrtacine® New Generation on P. acnes biofilm alone or combined with antibiotics was demonstrated and can lead to consider it as a potent adjunctive product efficient during the antibiotic course for acne vulgaris treatment.

International audience; (Pt-€“Pt-Ag/AgCl) and (Au-€“Pt-€“Ag/AgCl) electrochemical microcells (ElecCell) were developed for the detection of redox species by cyclic voltammetry. A special emphasis was placed on the SU-8 waferlevel passivation process in order to optimize the electrochemical properties of the different “thin film” metallic layers, i.e. gold or platinum for the working electrode, platinum for the counter electrode and silver/silver chloride for the reference electrode. (Au–Pt–Ag/AgCl) microcells were applied for the detection of antioxidant species such as ascorbic and uric acids in phosphate buffer solution, evidencing high sensitivity but low selectivity. Works were extended to skin analysis, demonstrating that a good electrical contact with the skin hydrolipidic film allowed the effective evaluation of the skin global antioxidant capacity.

Oxidative stress is associated to the massive generation of reactive oxygen species inducing fast oxidative reactions in chain. To overcome this problem, reasonable supplementations of antioxidants are widely practiced, mostly based on empirical protocols. An electrochemical process is proposed to choose the best molecules association inducing the greatest antioxidant capacity, by coupling homogeneous or heterogeneous catalytic reactions involving antioxidants with an electrochemical step. Cyclic voltammetry and constant potential electrolysis experiments were used to highlight regeneration reactions induced by specific associations of hydrophilic and lipophilic antioxidants in homogeneous medium or at liquid/liquid interface. Results showed that N-Acetyl-L-Cysteine (NAC) was effectively regenerated by both ascorbic acid (AA) and ascorbyl glucoside (AA-2G) but not by ascorbyl phosphate magnesium (AA-2P). The antioxidant properties of alpha-tocopherol also increased to 40% when associated with AA. The process was successfully applied for the first time to the study of simple emulsions. Formulations involving NAC and AA-2G presented the highest synergic effect, the antioxidant capacity being amplified by more than 35%. On the other hand no catalytic mechanism was observed when introducing three antioxidants in the cream because it induced too low reaction kinetics. This electrochemical process can thus be exploited as a tool for the optimisation of “anti-aging” dermocosmetic formulations.

Starting from a mixture of enantiomers in solution, crystallization can generate different types of crystals. In order to determine which type of crystal is obtained in the case of acetyl leucine, an active pharmaceutical ingredient (API), analytical methods have been used to partially elucidate the binary and ternary phase diagrams of the system composed of the two enantiomers and water. The melting temperature phase diagram of this compound has been obtained by using differential scanning calorimetry (DSC) analyzes. The results show that it is characteristic of a conglomerate. This mode of crystallization has also been confirmed by X-ray powder diffraction analysis. Solubility measurements of enantiomerical mixtures in water enabled the determination of the ternary diagram of solubility. The empiric Meyerhoffer double solubility rule has been modified, due to the characterization of interactions between enantiomers.

In pharmaceutical industries, active pharmaceutical ingredients (API) are made of crystals whose properties must be controlled because they influence the end-use properties of the drug. Even if crystal quality is mainly determined during the precipitation step, downstream processing also has an influence. In this study, the influence of washing on the crystal size and shape was analyzed. For the API being considered, different impurities have to be removed from the final suspension by filter cake washing. The efficiency of the washing steps was measured by different types of characterization on the solid phase (differential scanning calorimetry, scanning electron microscopy, and size distribution) and on the remaining filtrate (concentration of impurities). A second component also coprecipitates with the API. A specific study has been carried out on the withdrawal of this by-product and on its impact on the evolution of the crystalline form during washing steps. It was found that three filter cake washings allow us to remove all the impurities and to obtain a pure crystalline form.

Solubility, which defines the liquid /solid equilibrium, is a key parameter to control a crystallization process. This work is focused on the effects of pH, temperature and impurities on the aqueous solubility of an Active Pharmaceutical Ingredient (API).As the API is a weak acid (pKa = 3.7), its solubility increases with the pH. On the basis of the experimental curve of solubility, a model was defined to fit the evolution of the solubility as a function of pH. In the case of this compound, studies revealed a weak influence of the temperature in comparison with the pH. So, the solubility of the compound is slightly impacted by the temperature.Some experiments were carried out in order to compare the solubility of the API, at the same pH and temperature, for different concentrations of impurities found in the process. The results revealed a solubility increase in presence of acetic acid and a high solubility decrease in presence of sodium chloride. By carrying out experiments on common ions salts, the anion chloride Cl- has been identified as the cause of the solubility decrease.

Transposition to continuous and process intensification of a pharmaceutical production.

The electrochemical behaviour of N-acetyl-l-cysteine (NAC) has been investigated by linear and cyclic voltammetry on gold electrode at room temperature. The results showed two oxidation peaks under acid and neutral conditions and only one in basic medium. For each oxidation, as many electron was exchanged as proton. The influence of both the concentration and the potential scan rate on the peak currents highlighted a diffusion-controlled phenomenon for the first peak and an adsorption-limited reaction rate for the second one. The diffusion coefficient of NAC in solution and the surface concentration of the adsorbed species at pH 3 and 7 were close to 2 × 10−4 to 2 × 10−5 cm2 s−1 and 6 × 10−9 to 6 × 10−10 mol cm−2, respectively. Film transfer experiments resulted in an irreversible adsorption of NAC on gold electrode, and the formation of a self-assembled monolayer (SAM).

Cyclic voltammetry is proposed as a new method for evaluating the antioxidant capacity of skin based on the reducing properties of low molecular weight antioxidants (LMWA). Experiments were performed simply by recording the anodic current at 0.9 V/SCE of a platinum microelectrode placed directly on the epidermis surface without any gel or water. This method ensured a direct, rapid (less than 1 min), reliable (accuracy 12%) and non-invasive measurement of the global antioxidant capacity of the stratum corneum with a high spatiotemporal resolution. At the same time, the pH of the skin surface was determined by recording the cathodic current at 0 V/SCE. Based on an exploratory study involving nine volunteer subjects, the evolution of the amperometric response of the microelectrode with time revealed a periodic modification of the redox properties.